ABSTRACT: Blubber biopsy samples from eastern North Pacific killer whales Orcinus orca were analyzed for fatty acids, carbon and nitrogen stable isotopes and organochlorine contaminants. Fatty acid profiles were sufficiently distinct among the 3 reported ecotypes (resident, transient or offshore) to enable individual animals to be correctly classified by ecotype and also by mitochondrial DNA (mtDNA) haplotype. Profiles of PCBs also enabled unambiguous classification of all 3 killer whale ecotypes, but stable isotope values lacked sufficient resolution. Fatty acid, stable isotope and PCB profiles of the resident and transient ecotypes were consistent with those expected for these whales based on their reported dietary preferences (fish for resident whales, marine mammals for transients). In addition, these ecotype profiles exhibited broad similarity across geographical regions, suggesting that the dietary specialization reported for resident and transient whales in the well-studied eastern North Pacific populations also extends to the less-studied killer whales in the western Gulf of Alaska and Aleutian Islands. Killer whales of the same ecotype were also grouped by region of sample collection. The mean stable isotope ratios of various regional groups differed considerably, suggesting that the prey preferences of these North Pacific killer whales may be both region and ecotype specific. Furthermore, 3 specific ecotypes of killer whales were found to have measured stable isotope values that were consistent with dietary preferences reported in the literature. Finally, although the offshore population had blubber fatty acid profiles implicating fish as its primary prey, contaminant and stable isotope results were equally congruent with predation on marine mammals.